scholarly journals Gene structure and functional analysis of the human Na+/phosphate co-transporter

1997 ◽  
Vol 324 (3) ◽  
pp. 927-934 ◽  
Author(s):  
Yutaka TAKETANI ◽  
Ken-ichi MIYAMOTO ◽  
Keiko TANAKA ◽  
Kanako KATAI ◽  
Mika CHIKAMORI ◽  
...  
Keyword(s):  
Vitamin D ◽  
Vitamin D Receptor ◽  
Transcription Initiation ◽  
Thyroid Hormone Receptor ◽  
Direct Repeat ◽  
Initiation Site ◽  
Transcription Initiation Site ◽  
Receptor Expression ◽  
Specific Gene ◽  
Flanking Region

Three λ phage clones encompassing the Na+/phosphate co-transporter (NaPi-3) gene and its 5′ flanking region were isolated from a human genomic DNA library. The gene comprises 13 exons and 12 introns and spans approx. 14 kb. All exon–intron junctions conform to the GT/AG rule. The major transcription-initiation site was determined by primer-extension analysis and is an adenosine residue 57 bp upstream of the 3′ end of the first exon. There is a typical TATA box 28 bp upstream of the major transcription-initiation site and various cis-acting elements, including a cAMP-responsive element, AP-1, AP-2 and SP-1 sites in the 5′ flanking region. This region also contains three direct-repeat-like sequences that resemble the consensus binding sequence for members of the steroid–thyroid hormone receptor superfamily, including vitamin D. Deletion analysis suggests that the region from nt-2409 to nt-1259 in the 5′ flanking region may be involved in kidney-specific gene expression. Vitamin D responsiveness of the NaPi-3 promoter was also detected in COS-7 cells co-transfected with a human vitamin D receptor expression vector. The presence of the three vitamin D receptor-responsive elements in the NaPi-3 promoter may be important in mediating the enhanced expression of the gene by 1,25-dihydroxyvitamin D3.

scholarly journals Genomic footprinting of a yeast tRNA gene reveals stable complexes over the 5'-flanking region.

1989 ◽  
Vol 9 (8) ◽  
pp. 3244-3252 ◽  
Author(s):  
J M Huibregtse ◽  
D R Engelke
Keyword(s):  
Stationary Phase ◽  
Transcription Initiation ◽  
Initiation Site ◽  
Dnase I ◽  
Transcription Initiation Site ◽  
Upstream Region ◽  
Stable Complex ◽  
Coding Region ◽  
Flanking Region ◽  
Genomic Footprinting

We have shown by genomic footprinting that the 5'-flanking region of the Saccharomyces cerevisiae tRNASUP53 gene is protected from DNase I digestion. The protected region has a 5' boundary at -40 (relative to the transcription initiation site) and extends into the coding region of the gene, with a 3' boundary at approximately +15. Although the DNase I protection over this region was much greater than at the A- and B-box internal promoters, point mutations within the A or B box that reduced transcription in vitro eliminated the upstream DNase I protection. This implies that formation of a stable complex over the 5'-flanking region is dependent on interaction of the gene with transcription factor IIIC but that stability of the complex may not require continued interaction with this factor. The DNase I protection under varied growth conditions further suggested that the upstream complex is composed of two or more components. The region over the transcription initiation site (approximately +15 to -10) was less protected in stationary-phase cultures, whereas the more upstream region (approximately -10 to -40) was protected in both exponential- and stationary-phase cultures.

Genomic footprinting of a yeast tRNA gene reveals stable complexes over the 5'-flanking region

1989 ◽  
Vol 9 (8) ◽  
pp. 3244-3252
Author(s):  
J M Huibregtse ◽  
D R Engelke
Keyword(s):  
Stationary Phase ◽  
Transcription Initiation ◽  
Initiation Site ◽  
Dnase I ◽  
Transcription Initiation Site ◽  
Upstream Region ◽  
Stable Complex ◽  
Coding Region ◽  
Flanking Region ◽  
Genomic Footprinting

We have shown by genomic footprinting that the 5'-flanking region of the Saccharomyces cerevisiae tRNASUP53 gene is protected from DNase I digestion. The protected region has a 5' boundary at -40 (relative to the transcription initiation site) and extends into the coding region of the gene, with a 3' boundary at approximately +15. Although the DNase I protection over this region was much greater than at the A- and B-box internal promoters, point mutations within the A or B box that reduced transcription in vitro eliminated the upstream DNase I protection. This implies that formation of a stable complex over the 5'-flanking region is dependent on interaction of the gene with transcription factor IIIC but that stability of the complex may not require continued interaction with this factor. The DNase I protection under varied growth conditions further suggested that the upstream complex is composed of two or more components. The region over the transcription initiation site (approximately +15 to -10) was less protected in stationary-phase cultures, whereas the more upstream region (approximately -10 to -40) was protected in both exponential- and stationary-phase cultures.

scholarly journals Isolation and characterization of the human diacylglycerol kinase gene

1993 ◽  
Vol 294 (2) ◽  
pp. 443-449 ◽  
Author(s):  
K Fujikawa ◽  
S Imai ◽  
F Sakane ◽  
H Kanoh
Keyword(s):  
Transcription Initiation ◽  
Diacylglycerol Kinase ◽  
Initiation Site ◽  
Transcription Initiation Site ◽  
Jurkat Cells ◽  
Upstream Region ◽  
Negative Element ◽  
Isolation And Characterization ◽  
Flanking Region ◽  
Nuclease Mapping

The 80 kDa diacylglycerol kinase (DGK) is abundantly expressed in oligodendrocytes and lymphocytes but not to a detectable extent in other cells such as neurons and hepatocytes. As an initial attempt to delineate the mechanism of the transcriptional control of the DGK gene, we have cloned from a human genomic library a 22 kb genomic fragment. The genomic clone consists of the 5′-flanking region and 17 exons coding for approx. 53% of the total exons of human DGK, including those encoding EF-hand and zinc-finger regions. The translation initiation site is located in the second exon. S1 nuclease mapping and primer extension analysis of the human DGK mRNA identified a major transcription initiation site (position +1) at 264 bp upstream from the initiator ATG. In the 5′-flanking sequence we detected a single GC box at -35 but no canonical TATA and CAAT sequences. However, the sequence starting from the cap site (AGTTCCTGCCA) is very similar to the initiator element that specifies the transcription initiation site of some housekeeping genes. In addition, the 5′-upstream region contains several putative cis-elements. Jurkat and HepG2 cells were transfected with various 5′-deletion mutants of the upstream region fused to the structural gene of chloramphenicol acetyltransferase (CAT). The CAT assay revealed that among constructs containing up to 3.4 kb of the 5′-flanking region, a fragment of 263 bp from the transcription initiation site contains a basic promoter that is active in both types of cells. Moreover, the region between -263 and -850 contains a negative element that is active in HepG2 but not in Jurkat cells. This negative element may, at least in part, be responsible for the cell type-specific expression of the DGK gene.

scholarly journals Initiation binding repressor, a factor that binds to the transcription initiation site of the histone h5 gene, is a glycosylated member of a family of cell growth regulators [corrected].

1995 ◽  
Vol 15 (12) ◽  
pp. 6670-6685 ◽  
Author(s):  
A Gómez-Cuadrado ◽  
M Martín ◽  
M Noël ◽  
A Ruiz-Carrillo
Keyword(s):  
Dna Binding ◽  
Transcription Initiation ◽  
Direct Repeat ◽  
Initiation Site ◽  
Transcription Initiation Site ◽  
Nuclear Targeting ◽  
Dimerization Domain ◽  
Localization Signal ◽  
Molecular Aspects ◽  
Histone H5

Initiation binding repressor [corrected] (IBR) is a chicken erythrocyte factor (apparent molecular mass, 70 to 73 kDa) that binds to the sequences spanning the transcription initiation site of the histone h5 gene, repressing its transcription. A variety of other cells, including transformed erythroid precursors, do not have IBR but a factor referred to as IBF (68 to 70 kDa) that recognizes the same IBR sites. We have cloned the IBR cDNA and studied the relationship of IBR and IBF. IBR is a 503-amino-acid-long acidic protein which is 99.0% identical to the recently reported human NRF-1/alpha-Pal factor and highly related to the invertebrate transcription factors P3A2 and erected wing gene product (EWG). We present evidence that IBR and IBF are most likely identical proteins, differing in their degree of glycosylation. We have analyzed several molecular aspects of IBR/F and shown that the factor associates as stable homodimers and that the dimer is the relevant DNA-binding species. The evolutionarily conserved N-terminal half of IBR/F harbors the DNA-binding/dimerization domain (outer limits, 127 to 283), one or several casein kinase II sites (37 to 67), and a bipartite nuclear localization signal (89 to 106) which appears to be necessary for nuclear targeting. Binding site selection revealed that the alternating RCGCRYGCGY consensus constitutes high-affinity IBR/F binding sites and that the direct-repeat palindrome TGCGCATGCGCA is the optimal site. A survey of genes potentially regulated by this family of factors primarily revealed genes involved in growth-related metabolism.

P12. Vitamin D receptor expression and monocyte differentiation

Bone ◽  
1992 ◽  
Vol 13 (3) ◽  
pp. 278
Author(s):  
M. Hewison ◽  
R. Walker ◽  
J.L.H. O'Riordan
Keyword(s):  
Vitamin D ◽  
Vitamin D Receptor ◽  
Receptor Expression ◽  
Monocyte Differentiation
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